Adv. Hort. Sci., 2014 28(4): 214-224

Wild grape germplasms in

H. Yamashita*, R. Mochioka** * Faculty of Life and Environmental Sciences, University of Yamanashi, 4-4-37, Takeda, Kofu, Yamanashi, Japan. ** Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, Kita, Kagawa, Japan.

Key words: anthocyanins, breeding, classification, geographic distribution, growth cycle, .

Abstract: In Japan, seven species and eight varieties of wild grapes were identified, among which the main species are Vi- tis coignetiae Pulliat, V. flexuosa Thunb., and V. ficifolia Bunge var. lobata (Regel) Nakai (syn. V. thunbergii Sieb. et Zucc.). This paper summarizes the identification and classification of wild grapes native to Japan based on the past reports. Their distributions in Japan and physiological and ecological traits are also reviewed for effective practical use for grape breeding in the future.

1. Introduction cies found in Japan. Many other species exist locally in limited areas. In addition, researchers from Osaka Pre- It is thought that ancestors of grape (genus Vitis) ap- fecture University discovered Shiohitashibudou (tenta- peared during the first half of the Cretaceous period. They tive name) (Nakagawa et al., 1991). The geographical then spread around the world according to environmental distribution of the wild grapes native to Japan are shown and anthropogenic influences, and now comprise three ma- in figures 1-4. These figures were created from a site jor groups of species: European, North American, and East survey from to Okinawa starting in 1973, and Asian species, which differ in their physiological and eco- were made based on past records and reports using con- logical characteristics (Horiuchi and Matsui, 1996). Wild served (pressed) leaf specimens from Hokkaido Univer- grapes native to Japan belong to the group comprising the sity, Tokyo Metropolitan University, Kyoto University, East Asian species. Only a few reports on wild grape spe- Niigata University, Kumamoto University (Japan), and cies, including classification, physiological, and ecological Taiwan University (Taiwan). characterizations, have been published so far (Horiuchi and Matsui, 1996). Nevertheless, the importance of wild grapes Yamabudou, Vitis coingnetiae Pulliat (Fig. 1) as genetic resource for grape breeding has gradually been This species is widely distributed from level ground recognized because some wild grapes show superior traits to the lowest mountain areas in Hokkaido; from the low- towards global warming in terms of sustainable berry pro- est areas in the mountains to the mountain zone in the duction under hot and humid conditions. Tohoku district (northeastern region of Japan); from the This paper describes the identification and classifica- mountain zone to the alpine region in the Chubu district tion of wild grapes native to Japan. Their physiological (central region of Japan); and in the alpine regions of the and ecological traits, as well as their utilization are also Kinki, Chugoku, and Shikoku districts. It is thought that reviewed by focusing on the latest research findings re- this species is also present in a limited area of the alpine garding wild grapes native to Japan. regions in the Kyushu district, but it has not yet been dis- covered around Mt. Aso, which is consistent with the fact that we could not find any pressed leaf specimen in the 2. Geographical distribution universities located in the Kyushu distinct. It is notewor- thy that this species is not distributed in South , This, and previous studies, found that seven Vitis spe- China, and neighboring countries; including Far Eastern cies and eight varieties are distributed throughout Japan, Russia. However, it has been confirmed that Yamabudou from Hokkaido (northern region) to Okinawa (southern grows naturally in the South Chishima and Sakhalin dis- region) (Table 1) (Nakagawa et al., 1991). Of these, Yam- tricts (Horikawa, 1972). abudo, Ebizuru, and Sankakuzuru are the three main spe- Sankakuzuru (Gyojanomizu), V. flexuosa Thunb. (Fig. 2) Received for publication 17 September 2014 This species is distributed in the mid regions of the Accepted for publication 14 November 2014 Yamabudou (Fig. 1) and Ebizuru (Fig. 3) ranges, overlap-

214 Table 1 - Systematic and geographical distribution of wild grapes native to Japan (Nakagawa et al., 1991)

Species or varieties Japanese name Locality where grown Vitis coignetiae Pulliat Yamabudou Hokkaido, Honshu, Shikoku Vitis coignetiae Pulliat var. glabrescens Hara Takeshimayamabudou Hokkaido, Honshu Vitis flexuosa Thunb. Sankakuzuru (Gyojanomizu) Honshu, Shikoku, Kyusyu Vitis flexuosa Thunb. var. rufo-tomentosa Makino Kesankakuzuru Southern Honshu, Shikoku Vitis flexuosa Thunb. var. tsukubana Makino Usugesankakuzuru Northern Honshu Vitis flexuosa Thunb. var. crassifolia Hara Atsubasankakuzuru Shikoku Vitis saccharifera Makino Amazuru (Otokobudou) Southern Honshu, Shikoku Vitis yokogurana Makino Yokogurabudou Shikoku (Kochi Pref.) Vitis ficifolia Bunge var. lobata (Regel) Nakai Ebizuru (z) All over Japan (Vitis thunbergii Sieb. et Zucc.) Vitis ficifolia Bunge var. izu-insularis Hara Shititouebizuru Izu Islands Vitis ficifolia Bunge var. sinuata Hara Kikubaebizuru Southern Honshu, Shikoku, Kyusyu Vitis ficifolia Bunge var. ganebu Hatusima Ryuukyuuganebu Amami Islands, Okinawa Islands, Yaeyama Islands Vitis austrokoreana Hatusima Kenashiebizuru Tsushima Islands Vitis kiusiana Momiyama Kumagawabudou Kyushyu (Kumamoto pref., Kagoshima pref.) Vitis shiragai Makino Shiragabudou Honshu (Okayama pref.) Vitis sp. Shiohitashibudou (tentative) Kyusyu (Kagoshima pref.) (z) used in some classifications as a species (thunbergii).

Fig. 1 - Geographic distribution of Vitis coignetiae Pulliat (Nakagawa Fig. 2 - Geographic distribution of Vitis flexuosa Thunb. (Nakagawa et et al., 1986). al., 1986).

215 ping with the two species, and is found in slightly lower al- of Japan. Takeshimayamabudou, a variety of Yamabudou titude areas than Yamabudou (Fig. 1). We can usually find (V. coingnetiae), was discovered in Hokkaido (around this species from the lowlands to the mountainous area of Lake Akan) and Nagano prefecture. It has no hairiness on the Tohoku district or the Chubu district; it does not grow the lower leaf surface and a thinner leaf compared to Yam- naturally in Hokkaido. abudou. Kumagawabudou has prickly shoots and ovoid leaves; one wild grape (V. davidii) with prickly shoots Ebizuru, V. ficifolia Bunge var. lobata (Regel) Nakai (Fig. 3) grows naturally in China, however, it differs significantly This species is one of the most widespread Vitis species from Kumagawabudou in its morphological characteris- in Japan. Its distribution extends from the southern Hok- tics. Shichitouebizuru grows in seaside areas of seven is- kaido region to the flatlands and mountainous terrain in lands of Izu. Moreover, its fruit-set is the highest among the Okinawa district; it can be found in a wide variety of all wild grapes native to Japan. habitats, including both the seashore and urban districts. This species is considered to be highly adaptable to the environment, thus it has a wide distribution compared with 3. The classification of Japanese wild grapes other Vitis species. As a result, many variants of morpho- logical and physiological traits are found in this species as As genus Vitis is mainly classified by morphology, a result of adaptation to local climates. Ryuukyuuganebu, some taxa may be taken as different classifications even Shichitouebizuru, and Kikubaebizuru are varieties belong- if they are the same grape. For example, Shiragabudou ing to V. ficifolia. Kenashiebizuru is also closely related to has two scientific names, Vitis shiragai Makino (Makino, V. ficifolia, although its scientific name is given as V. aus- 1918) and Vitis amurensis Rupr. (Ohwi, 1953). In this sec- koreana Hatusima. These grapes are generally included in tion, wild grapes native to Japan are classified concisely the V. ficifolia group. based on chemical, biochemical and genetic knowledge.

Other species (Fig. 4) Morphological classifications As shown in Table 1 and figure 4, in addition to these, Classification by leaf structure. Galet (1979) tried to many species and varieties are spread in various districts classify genus Vitis through ampelographic measurements

Fig. 3 - Geographic distribution of Vitis ficifolia Bunge var. lobata (Re- Fig. 4 - Geographic distribution of wild grapes native to Japan (Nak- gel) Nakai (Nakagawa et al., 1986). agawa et al., 1986).

216 of the leaf (Fig. 5, Table 2). Nakagawa et al. (1991) coded Cordiform: 357 to 468, Cuneiform: 135 to 247, Truncate: the characteristics of grape leaves and the result is present- 045 to 247, Orbicular: 015 to 136, Reniform: 014 to 136. ed in Table 3. Code numbers of vein length rations (ABC) According to this method, code numbers are relatively for the five basic leaf shapes are as follows (Galet, 1979): near for close species (e.g. V. coignetiae and V. amurensis) (Table 3). Classification by pollen ultrastructure. Mochioka et al. (1993) observed ultrastructures of mature pollen grains of wild grapes native to Japan, Korea and China using a scanning electron microscope and reported that the pollen could be classified as one of three types by the lumina forms in muri (Fig. 6). They also reported the

Table 3 - The code number of various wild grapes obtained by using the Galet’s method (Nakagawa et al., 1991)

Species ABC-r-S1S2 Vitis coignetiae 146-4-24 Vitis amurensis 146-3-24 Vitis flexuosa 357-7-01 Vitis ficifolia var. lobata 246-3-13 Vitis shiragai 136-3-13 Vitis sp. (Daisankakuzuru) 146-3-12 Vitis ficifolia Bunge var. ganebu 135-3-24 Vitis sp. (Shiohitashibudo) 257-5-02 Vitis ficifolia var. izu-insularis 146-3-01 Vitis kiusiana 368-7-01

Fig. 5 - The general morphology of a mature grape leaf. al.= Apical lobe; II= Lateral lobe; pl.= Proximal lobe; ss.= Superior sinus; is.= inferior sinus; ps. Petiolar sinus; pe.= Petiol; mv.= Mid- vein; sv.= Superior lateral vein; iv.= Inferior lateral vein; pv.= petiolar vein; vl.= Veinlet; se.= Serration; lm= Leaf margin. (Nakagawa et al., 1991).

Table 2 - The code number of the Galet (1979) ruler for the values of

A, B, C, r, S1 and S2 Code Vales of A, B and C Vales of r Vales of S Values of S number 1 2 0 0.91~1.00 ≤0.80 ≤ 70° ≤ 100° 1 0.81~0.90 0.81~0.90 71°~80° 101°~110° 2 0.71~0.80 0.91~1.00 81°~90° 111°~120° 3 0.61~0.70 1.01~1.10 91°~100° 121°~130° 4 051~0.60 1.11~1.20 101°~110° 131°~140° 5 0.41~0.50 1.21~1.30 111°~120° 141°~150° 6 0.31~0.40 1.31~1.40 121°~130° 151°~160° 7 0.21~0.30 1.41~1.50 131°~140° 161°~170° 8 0.11~0.20 141°~150° 171°~180° 9 0.00~0.10 ≥151° ≥181°

A= L2 length/L1 length; B= L3 length/L1 length; C= L4 length/L1 length, where L1= Midwein; L2= Superior lateral vein and L3= Inferior lateral vein; L4= Petiolar vein. Fig. 6 - Scanning electron microphotographs of grape exine. Scale bars= r= Leaf length/leaf width. 1 µm. Type-1 A pollen has perforations in the lumina; type-1 B

S1= α + β; S2= α + β + r where α= Angles between L1 and L2. β= Angles pollen has perforations and corrugation in the lumina, and type-II between L2 and L3. and r= Angles between L3 and L4. pollen has granules in the lumina (Mochioka et al., 1993).

217 pollen ultrastructures of related species belonged to the et al. (1995) analyzed by HPLC anthocyanins in the berry same type (Table 4). skin of 10 wild grapes (four species, five varieties and one unidentified type) native to Japan. The dendrogram, show- Chemotaxonomic classifications ing phylogenetic relationships, was drawn from the pairwise Classification by anthocyanins in grape skin. Mochioka comparison of matching coefficients based on anthocyanin

Table 4 - Morphological characteristics of pollen grains of the wild grapes native to Japan, Korea and China (Mochioka et al., 1993)

Pollen size (µm) No. of Pollen Species or cultivars L/W ratio Length (L) Width (W) colpi exine type Japan Vitis coignetiae Pulliat ♂ 20.0±0.2 (z) 19.4±0.2 1.03±0.01 3 I A ♀ 22.0±0.2 20.7±0.2 1.06±0.01 0 I A Vitis flexuosa Thunb. ♂ 20.3±0.5 19.6±0.3 1.04±0.03 3 I A ♀ 20.6±0.2 20.1±0.2 1.03±0.01 0 I B, II Vitis ficifolia Bunge var. lobata (Regel) Nakai ♂ 20.0±0.2 19.2±0.2 1.04±0.01 3 II ♀ 20.9±0.2 20.0±0.2 1.05±0.01 0 II Vitis ficifolia Bunge var. izu-insularis Hara ♂ 20.9±0.3 20.1±0.4 1.04±0.01 3 II ♀ 21.4±0.2 20.2±0.2 1.06±0.01 0 II Vitis ficifolia Bunge var. ganebu Hatusima ♀ 21.4±0.3 20.3±0.2 1.05±0.01 0 I A Vitis shiragai Makino ♂ 21.1±0.2 20.7±0.2 1.02±0.01 3 I A ♀ 22.6±0.2 21.1±0.3 1.08±0.01 0 I A Vitis kiusiana Momiyama ♀ 20.4±0.2 19.5±0.2 1.05±0.01 0 I B Vitis sp. (provisional name: Shiohitashibudo) ♀ 21.3±0.3 20.0±0.3 1.06±0.01 0 II Korea Vitis amurensis Rupr. ♂ 20.6±0.3 20.1±0.2 1.03±0.01 3 I A ♀ 23.4±0.2 22.1±0.2 1.06±0.01 0 I B Vitis sp. (provisional name: Daisankakuzuru) ♀ 21.4±0.2 20.3±0.2 1.06±0.01 0 I B China Vitis amurensis Rupr. ♂♀ 21.5±0.2 20.7±0.2 1.04±0.01 3 I B Vitis flexuosa Thunb. ♂ 19.8±0.3 19.0±0.3 1.04±0.01 3 I A Vitis ficifolia Bunge ♂ 20.6±0.3 19.0±0.3 1.04±0.01 3 II Vitis adstricta Hance ♂ 21.5±0.2 21.2±0.2 1.02±0.01 3 II Vitis adstricta Hance var. ternata W.T. Wang ♂ 22.3±0.3 21.0±0.4 1.06±0.01 3 II Vitis bellula (Rehd.) W.T. Wang ♂ 19.3±0.3 18.8±0.2 1.03±0.01 3 II Vitis davidii (Roman.) Foëx ♂ 23.1±0.2 22.4±0.2 1.03±0.01 3 I B ♂♀ 20.4±0.2 19.9±0.1 1.03±0.01 3 I B Vitis pseudoreticulata W.T. Wang ♂ 21.3±0.2 20.8±0.2 1.03±0.01 3 I A Vitis hancokii Hance ♂ 20.1±0.2 19.6±0.1 1.03±0.01 3 I B Vitis chugii Metcalf. ♂ 19.9±0.2 19.3±0.2 1.03±0.01 3 I B Vitis chunganensis Hu ♂ 19.9±0.2 19.7±0.2 1.01±0.01 3 I A Cultivar L. ‘Muscat of Alexandria’ ♂♀ 23.2±0.4 22.4±0.4 1.03±0.01 3 I B Vitis lambrusca L. ‘Concord’ ♂♀ 23.3±0.3 22.9±0.3 1.02±0.01 3 I B Vitis lambruscana Bailey ‘Delaware’ ♂♀ 21.3±0.3 20.1±0.3 1.06±0.01 3 I B ‘Campbell Early’ ♂♀ 24.6±0.4 23.4±0.4 1.05±0.01 3 II ‘Kyoho’ (tetraploid) ♂♀ 28.2±0.3 26.9±0.3 1.05±0.01 3.4 I A (z) Each valure represents the mean of 20 individual measurements ±se.

218 components, and agrees well with the morphological tax- of Pgm-2 existed only in wild grapes native to East Asia onomy (Fig. 7, Table 5) (Mochioka et al., 1995). (Table 6). In this study 19 anthocyanins were identified, and While restriction fragment length polymorphism there were more kinds of anthocyanins in the berry skins (RFLP) and random amplified polymorphic DNA of wild grapes distributed in southern regions than those (RAPD) analyses were used to analyze the relationships of wild grapes distributed in northern regions (Table 5). among wild and cultivated grapes, a phenogram of RAPD Classification by isozyme and DNA analysis. Species- data obtained showed a clear separation between wild specificity was observed in the alleles dominated by Gpi- and cultivated grapes (Goto-Yamamoto et al., 1998). 2 and Pgm-2 gene loci (Fig. 8, Table 6) (Ohmi et al., 1991). The F band of Gpi-2 and the A and the C bands

Fig. 7 - Dendrogram of berry skin anthocyanin phenotypes of wild and Fig. 8 - GPI and PGM isozymes bands coded by each alleles at Gp1-2 cultivated varieties (Mochioka et al., 1995). (left) and Pgm-2 (right) loci (Ohmi et al., 1991).

Table 5 - Percentage of anthocyanin composition in grape berry skins analyzed by HPLC (z) (Mochioka et al., 1995)

Peak No (y) Species or cultivars 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Wild grape Vitis coignetiae 1 4 66 1 4 2 1 18 2 V. amurensis 2 2 81 V. flexuosa 5 4 4 57 7 4 1 12 2 V. flexuosa var. rufo-tomentosa 2 1 28 3 3 2 1 4 1 38 2 8 V. ficifolia var. lobata 6 2 10 6 44 2 2 1 7 5 10 12 1 V. ficifolia var. izu-insularis 15 3 17 5 27 3 1 3 12 5 2 2 4 V. ficifolia var. ganebu 8 4 9 8 10 28 2 4 3 4 1 3 2 9 1 V. ficifolia var. sinuata 10 5 39 13 8 9 4 3 6 1 2 1 V. shiragai 11 7 5 13 15 26 2 1 3 4 2 2 2 4 V. kiusiana 13 10 28 19 11 1 1 6 3 3 2 1 V. sp. (tentative name: Shiohitashibudou) 6 3 2 10 15 37 2 6 1 1 4 1 1 2 2 V. sp. (tentative name: Daisankakuzuru) 5 3 28 17 12 7 7 4 3 2 3 2 2 3 V. sp. (tentative name: Taiwan kes- 5 2 24 6 36 1 8 3 3 2 2 2 3 ankakuzuru) Cultivars ‘Delaware’ 44 26 4 3 9 4 ‘Muscat Bailey A’ 3 21 4 32 23 2 2 1 2 2 1 2 ‘Campbell Early’ 6 6 12 46 6 2 7 6 (z) Absorbance at 520 nm. (y) Peak No. 3= delphinidia 3-monoglucoside, no. 5= cyaniding 3-monoglucoside; no. 9= petunidin 3-monoglucoside; no. 16= malvidin 3-monoglucoside.

219 Table 6 - Species-specific alleles at 2 loci in grape (Ohmi et al., 1991) Furthermore, ecological differences exist between Shi- ragabudou and V. amurensis. Shiragabudou is distributed Locus Allele Species (z) over the warm lowland from 20 to 240 m above sea level Gpi-2 A Vin. in Okayama while V. amurensis has a growth area in the B Vin., Amur. (?) cold districts at 40 to 50° N latitude. The cross section C Lab., Rip. (?) morphology of Shiragabudou shoots is hexagonal, while D Vin. that of V. amurensis is circular. E Lab., Shir. These differences show that Shiragabudou and V. F Thun., Shir. amurensis are not the same species, thus Vitis shiragai Makino should be used as the scientific name for Shira- G Vin., Amur. gabudou. H Lab., Aest. (?), Vulp (?) Ebizuru and its varieties. Ebizuru is distributed wide- I Lab., Rup., Champ., Linc. (?), Coig., ly in Japan. There are a number of varieties and ecotypes Shir. in the Ebizuru group, and morphological differences are J Bourq. and/or Lab. various. Even if they are the same species, there are sev- K Champ. eral synonyms for this group. Even now, Vitis ficilolia, V. L Vulp. and/or Lab. ficifolia var. lobata, and V. thunbergii are used as scientif- ic names for Ebizuru. An isotype of V. ficifolia is the wild Pgm-2 A Shir. grape native to China. There are definitely differences in leaf morphology and bearing habit between Chinese fici- B Vin. folia and Japanese Ebizuru. C Amur., Coig., Thun., Shir. Natural hybrids. Different Vitis species can be hy- D Vin. bridized easily with each other, so there are many natural E Vin. hybrids. V. yokogurana is supposed to be a hybrid of V. F Lab. and/or Linc. flexuosa and V. saccharifera (Makino, 1918); V. flexuosa G Lab., Aest. (?) var. tukubana is supposed to be a hybrid of V. flexuosa H Vin., Rup., Champ., Lab., Coig., Aest. and V. ficifolia var. lobata (Murata, 1971). (?), Bourq. (?), Rip. (?), Vulp. (?) Yamabudou. Yamabudou (V. coignetiae) must be a I Vin. species related to V. amurensis, but there is no report that both these two species are simultaneously distributed J Rup. over the same regions in Japan or other countries. K Champ. Shiohitashibudou. (tentative name): Shiohitashibu- (z) Abbreviations: Amur.= Vitis amurensis; Aest.= V. aestivalis; Bourq.= dou is an unidentified species, and it was discovered in V. aestivalis var. bourquiana; Champ.= V. champini; Lab.= V. labrusca; Kagoshima prefecture, Kyushu (Nakagawa et al., 1991). Linc.= V. lincecumii; Rip.= V. riparia; Rup.= V. rupestris; Shir.= V. shi- Its leaf morphology is definitely different from that of ragai; Tun.= V. thunbergii (=V. ficifolia var. lobata); Vin.= V. vinifera; Vulp.= V. vulpina. other Japanese wild grapes. Shiohitashibudou might be a related species or natural hybrid of Ebizuru because its flowering habit is ever-bearing and the pollen ultra- structure is type II. Its bud endodormancy is deeper, and Classification by general judgments the soluble solid content of its juice is higher than other Since the past horticultural classification was Japanese wild grapes (Mochioka, 1996). qualitatively performed considering a small number of Ryuukyuuganebu. Ryuukyuuganebu is distributed characteristics, a different result for some researchers over Amami, Ryukyu and Yaeyama Islands, and is sup- might be found (e.g. Shiragabudou). Therefore, using plu- posed to be a variety of V. ficifolia. However its leaf shape ral classification methods is desirable because just one is different from V. ficifolia var. lobata (Fig. 9, Table 3), method might induce the wrong result. and its pollen ultrastructure is also different (Table 4). What follows is a brief discussion of judgments about Ryuukyuuganebu is ever-green in its habitat. some Japanese wild grapes with questionable taxonomic points. Shiragabudou. Shiragabudou was discovered in 4. Physiological and ecological traits of wild grapes na- Okayama prefecture, Honshu and was first named by tive to Japan Makino (1918). As the leaf shape of this wild grape re- sembles that of V. amurensis Rupr. Ohwi (1953) changed Four species [V. coignetiae, V. flexuosa, V. shiragai, and its scientific name to Vitis amurensis Rupr. The leaf Shiohitashibudou (tentative name. Vitis sp.)] and two va- morphology and pollen ultrastructures of these two wild rieties (V. ficifolia var. lobata and V. ficifolia var ganebu ) grapes are in the same group (Table 3, 4), but anthocyan- of wild grapes native to Japan, and two species [Chosen in composition in berry skins (Table 5) and species-spe- Yamabudou (V. amurensis) and Daisankakuzuru (tentative cific alleles at 2 loci (Table 6) are apparently different. name. V. sp.)] grown in Korea, and ‘Delaware’ (V. labrus-

220 Growth cycle Bud burst. Bud burst of Yamabudou and Chosen Yam- abudou occurred earlier among the wild grapes, followed by Sankakuzuru, Ebizuru, and Daisankakuzuru. Shiohi- tashibudou showed the latest bud burst in this study (Table 7). ‘Delaware’ broke bud later than all wild grapes except for Shiohitashibudou. Full bloom (50% cap off). In grape cultivation, early bud burst does not always mean early bloom. Indeed, the orders of bloom date differed from those of bud burst (from early to late bloom): Yamabudou, Sankakuzuru Ebi- zuru/Shiragabudou, Daisankakuzuru, Shiohitashibudou, and Ryuukyuuganebu. Here, Ebizuru blooms in the same period as Shiragabudou, and the bloom date of Sankaku- zuru was comparable to that of ‘Delaware’ (Table 7). Veraison (berry coloring begins). The skin color of Yamabudou changed earliest among the wild grapes. Changes in skin color in Chosen Yamabudou and ‘Dela- ware’ occurred on the same date. Skin color change oc- curred the latest in Daisankakuzuru, Shiohitashibudou, and Ryuukyuuganebu; and Sankakuzuru, Shiragabudou and Ebizuru were the next to latest (Table 7). Maturity. Maturity is denoted when berry weights and soluble solids attain maximum maturation. ‘Delaware’ matured in mid-August, which was earlier than the studied wild grapes. Sankakuzuru and Ryuukyuuganebu matured in mid-September and Daisankakuzuru in early-October (Table 7). Thus, wild grapes tend to have medium to late maturation in Osaka. Defoliation. Defoliation indicates the date when all leaves (from basal to tenth leaf) fall completely. Defo- Fig. 9 - Standard design of mature leaf of various wild grapes (├─┤ 1 liation in Sankakuzuru and Daisankakuzuru took place cm) (Nakagawa et al., 1991). in early-November, Yamabudou and Chosen Yamabudou shed their leaves in mid-November, and Shiohitashibudou, Ebizuru, and Shiragabudou in late-November. Interest- cana Bailey) (Table 7) were cultivated in the Horticultural ingly, Ryuukyuuganebu, a subtropical grape, showed ex- Experiment Field at Osaka Prefecture University (Sakai, tremely late defoliation; in some cases, leaves did not fall Osaka) and their physiological and ecological traits were until January. compared (Nakagawa et al., 1986). Here, Chosen Yamabu- dou is a wild grape grown naturally in the northern and Characteristics of organ northeastern regions of China, northern region of Korea Shoot. Observations of shoot growth in summer en- and southeastern region of the former Soviet Union. The abled us to classify the wild grapes into three types: con- results are illustrated below. tinuous, subcontinuous, and discontinuous. The continu-

Table 7 - Growth cycle of wild grapes native to Japan at Sakai Osaka (Nakagawa et al., 1986)

Species and varieties Bud burst Full bloom Veraison Harvest Leaf fall Chosen yamabudou (V. amurensis Rupr.) 3/27 5/14 8/1 9/29 11/14 Yamabudou (V. coignetiae Pulliat) 3/27 5/12 7/26 9/29 11/13 Sankakuzuru (V. flexuosa Thunb.) 3/29 5/26 8/6 9/14 11/7 Ebizuru [V. ficifolia Bunge var. lobata (Regel) Nakai] 3/29 6/4 8/11 9/11 11/27 Shiragabudou (V. shiragai Makino) 3/29 6/4 8/6 9/15 11/23 Daisankakuzuru (tentative Vitis sp.) 3/29 6/10 9/4 10/6 11/4 Shiohitashibudou (tentative Vitis sp.) 4/10 6/13 9/4 9/22 11/25 Ryuukyuuganebu (V. ficifolia Bunge var. ganebu Hatusima 4/3 6/19 8/31 9/20 after end of Dec. Delaware (V. labruscana Bailey) 4/6 5/27 7/30 8/20 11/8

221 ous group had constant growth of some shoots in summer; with two to four inflorescences at the basal part on each Shiohitashibudou and Daisankakuzuru were included in shoot without any inflorescences on lateral shoots. “B” this type. The subcontinuous type exhibited slight growth type: as seen in Shiohitashibudou, Shichitouebizuru, and of some shoots in summer; Ryuukyuuganebu, Sankakuzu- some Shiragabudou, there are two to four inflorescences at ru, Ebizuru, and Shiragabudou corresponded to this type. the basal part on each shoot with some inflorescences on Finally, the discontinuous group stopped shoot growth in lateral shoots. “C” type: Ebizuru and Ryuukyuuganebu be- summer, for example Yamabudou and Chosen Yamabudou long to this type, with two to six inflorescences at the basal in this study. Tendril placement of all Vitis species and va- part and the upper part of each shoot, having contiguous rieties native to Japan is intermittent. inflorescences from the base to the top of lateral shoots. Inflorescence. Wild grapes native to Japan (seven spe- cies and eight varieties) are dioecious, meaning that they Grape comparison and fruit quality (Table 8) contain imperfect individual male and female . Three Sugars. Shiohitashibudou attained a sugar concentra- types of fruiting habits were found (Fig. 10). “A” type: as is tion of 17.7%, which is the highest among the other spe- the case of Yamabudou, Chosen Yamabudou, Sankakuzu- cies (measured as 12-14%) except Ebizuru and Ryuukyuu- ru, Daisankakuzuru, Shiragabudou, and Kumagawabudou, ganebu, which had sugar concentrations of around 8%. Almost all studied species have high glucose content, especially Kumagawabudou, Shiohitashibudou, and Dai- sankakuzuru, which contained two to three fold more glu- cose than fructose. Acids. Kumagawabudou and Daisankakuzuru contained about 0.7 and 0.8% of organic acids, while almost all other species contained about 0.5%; Shiragabudou showed the lowest level of organic acids (0.36%) (Table 8). Amino acids. The concentration of amino acids var- ied widely from 50 mg% (Ebizuru) to 294 g% (Shiohi- tashibudou). Anthocyanins. Northern species such as Yamabudou and Chosen Yamabudou showed small amounts of antho- cyanin, while the southern species, Ryuukyuuganebu, con- tained a larger amount of this component (Table 8).

Dormancy and cold hardiness Dormancy. Although all species have a dormancy trait, its intensity depends on the species. Ryuukyuuganebu showed a short dormancy period, while it was generally longer in Kumagawabudou and Shiohitashibudou (Nak- agawa et al., 1986; Nakagawa, 1989). Cold hardiness. Cold hardiness varied markedly among the wild grapes. For example, Ryuukyuuganebu and Ku- magawabudou were very susceptible to severe damage at -3°C. On the contrary, Yamabudou, Ebizuru, Shiragabu- Fig. 10 - Fruiting habit of wild grapes native to Japan.

Table 8 - Berry composition of wild grapes native to Japan (Nakagawa et al., 1986) Reducing Glucose Fructose Glucose/ Organic Amino acid Anthocuanin Species and varieties sugars (%) (%) (%) Fructose acid (%) (mg %) (OD 537 nm) Chosen yamabudou (V. amurensis Rupr.) 12.7 9.2 3.5 2.6 0.50 191.2 0.12 Yamabudou (V. coignetiae Pulliat) 12.3 5.2 7.1 0.7 0.50 155.3 0.21 Sankakuzuru (V. flexuosa Thunb.) 14.2 8.0 6.2 1.3 0.52 214.0 0.30 Ebizuru [V. ficifolia Bunge var. lobata (Regel) Nakai] 8.1 4.1 4.0 1.0 0.51 50.7 0.30 Shiragabudou (V. shiragai Makino) 12.0 6.7 5.3 1.3 0.36 222.6 0.45 Daisankakuzuru (tentative Vitis sp.) 12.3 8.1 3.9 2.2 0.81 250.8 0.42 Kumagawabudou (V. kiusiana Momiyama) 12.0 9.2 2.8 3.3 0.72 180.5 0.49 Shiohitashibudou (tentative Vitis sp.) 17.7 12.7 5.0 2.5 0.48 294.5 0.41 Ryuukyuuganebu (V. ficifolia Bunge var. ganebu Hatusima 7.8 3.7 4.1 0.8 0.51 138.0 0.70 Delaware (V. labruscana Bailey) 16.8 7.5 9.3 0.8 0.77 220.8 0.05

222 dou, Shichitouebizuru, and Chosen Yamabudou showed iii) Salt tolerance moderate cold hardiness; their survival has even been re- Salt accumulation through the use of chemical fertilizers ported at -10°C (Nakagawa, 1989). causes serious problems which sometimes result in the loss of plants. Shiohitashibudou is excellent in its resistance to salts and Ryuukyuuganebu grows naturally along the sea- 5. Value and use of wild grape germplasms in Japan shore. Although the mechanism underlying salt tolerance of this grape has not yet been fully elucidated, this trait could Wine be useful not only for scion but also for root stock. Between the 1960s and 1980s, Yamabudou was suc- iv) Heat tolerance cessfully cultivated in commercial vineyards for wine- As global warming progresses, fruit skin, including making in the town of Ikeda (Hokkaido); its cultivation grape, shows poor coloration, which leads to a defective has attracted attention as a means to revitalize towns in appearance and reduced commercial value. Poor coloration Japan. In China, V. quinquangularis is processed into an also affects wine production. Interestingly, the coloration excellent wine (Li et al., 1992). Kumagawabudou, which of wild grapes native to Japan is very high, even under is thought to be the same species as V. quinquangularis (Li high temperature conditions during their ripening season. et al., 1991), therefore, may be an important resource for In grape, since MYB is involved in red skin coloration, it wine making. is valuable to compare MYB genes between wild grapes and primary Japanese cultivars such as ‘Kyoho’ and ‘Aki Breeding Queen’. Thus, this trait could be useful for sustainable The major cultivars, bred using wild grapes, are grape production with high quality and high adaptability ‘Sawanobori Waingurando’ and ‘Yama Sauvignon’ in Ja- in the production area. pan, both of which are used for wine production. ‘Sawa- nobori Waingurando’ is a cross seedling of V. amurensis × (Seibel 13053 × Nakajima No.1, a strain of Yamabu- 6. Conclusions dou) and it was released in 1998. By contrast, ‘Yama Sauvignon’ is a progeny of Yamabudou × ‘Cabernet Wild grapes native to Japan have been actively stud- Sauvignon’, which was released in 1990 by Dr. Yoshi- ied over a long period, but in recent years attention has hide Yamakawa at the University of Yamanashi. ‘Yama declined. However, wild grapes native to Japan can of- Sauvignon’ has the following superior characteristics: fer many useful characteristics, such as short dormancy, 1) no cracking of berry; 2) resistance to ripe rot, downy ever bearing, heat and salt tolerance. These traits are very mildew, and gray mold; 3) adaptability to the prevailing attractive, not only for their use as rootstock, but also in weather conditions in Japan; 4) high productivity; and terms of breeding material or for genetic studies. We have 5) high quality wine with typical aroma and taste (Ya- recently begun a breeding study using Ryuukyuuganebu. makawa et al., 1989). It is expected that some novel grapes will be bred to with- Considering the potential use of wild grapes as breeding stand increasing global temperatures or for use in grape material, these grapes have the following notable charac- cultivation factories. teristics: drought resistance, cold hardiness, salt tolerance, water logging tolerance, heat tolerance, disease resistance, high concentration of important substances, ever bearing, References and short dormancy. Four characteristics are especially promising: i) ever bearing, ii) short dormancy, iii) salt tol- GALET P., 1979 - A practical ampelography: Grapevine iden- erance, and iv) heat tolerance. Therefore, we explain the tification. - Cornell University Press, New York, USA, pp. usefulness of these characteristics for grape breeding pro- 24-47. grams in more detail. GOTO-YAMAMOTO N., MOCHIOKA R., LIN B., HASHI- i) Ever bearing ZUME K., UMEDA N., HORIUCHI S., 1998 - RFLP and Strains of Ryuukyuuganebu and Ebizuru bloom and RAPD analysis of wild and cultivated grapes (Vitis spp.). - J. fruit as long as the growth of axillary buds continues. This Japan. Soc. Hort. Sci., 67: 483-490. trait makes it possible to carry out year-round culture and/ HORIKAWA Y., 1972 - Yamabudo. Atlas of the Japanese flora: or culture using a factory system, like some vegetables, us- An introduction to plant sociology of East Asia. - Gakken ing artificial light and controlled irrigation. However, the Co., Ltd., Tokyo, Japan, pp. 208. major gene related to this trait has not yet been identified. HORIUCHI S., MATSUI H., 1996 - Nihon Budougaku. - Yok- ii) Short dormancy endo, Tokyo, Japan, pp. 58-102. Ryuukyuuganebu can be released from dormancy af- LI S.C., HORIUCHI S., MOCHIOKA R., MATSUI H., 1992 - ter being subjected to low temperature for extremely short Some issues on wild grapes native to East Asia (2). - Agric. periods. This trait may be profoundly related to ever bear- Hortic. (Nogyo-oyobi-Engei), 67: 1277-1280. ing. Thus, it may be possible to force culture inside green- LI S.C., HORIUCHI S., MOCHIOKA R., YAMAZAWA M., houses due to the low cost, for year-round culture and/or MATSUI H., 1991 - Comparison between wild grapes native culture using a factory system. to Japan and China. 2. Study on morphological character-

223 istics of V. kiusiana and V. quinquanglaris. - J. Japan. Soc. usage of gene resorces in fruit trees. - Report for Grants in Hort. Sci., 60(Suppl. 1): 132-133. Aid for Scientific Research, pp. 10-20. MAKINO T., 1918 - A contribution of the knowledge of the flora NAKAGAWA S., HORIUCHI S., MATSUI H., YUDA E., 1986 of Japan. - J. Jpn. Bot., 1: 31-33. - Studies on the cultivation and use of the wild grapes native MOCHIOKA R., 1996 - Taxonomic classification and utiliza- to Japan. - Report for Grants in Aid for Scientific Research, tion on wild grapes native to Japan based on their various pp. 1-49. characteristics. - Ph. D. Thesis, Osaka Prefec. University, NAKAGAWA S., HORIUCHI S., MATSUI H., YUDA E., YA- Sakai, Japan. MADA S., MURAI Y., KOMATSU H., 1991 - Distribution MOCHIOKA R., HORIUCHI S., LI S.C., MATSUI H., MURAI and leaf morphology of wild grapes Native to Japan. - J. Ja- Y., 1993 - Pollen morphology of wild grapes native to East- pan. Soc. Hort. Sci., 60: 31-39. ern Asia. - Jpn. J. Palynol., 39: 11-20. OHMI C., WAKANA A., SHIRAISHI S., 1991 - Phylogenic im- MOCHIOKA R., YAMAGUCHI M., HORIUCHIL S., MAT- plication of genus Vitis on the basis of isozyme analysis. - J. SUI H., KUROOKA H., 1995 - Chemotaxonomic classifica- Japan. Soc. Hort. Sci., 60(Suppl. 2): 62-63. tion of native wild Japanese grapes by anthocyanins in berry OHWI J., 1953 - Vitis Linn. Flora of Japan. - Shibundo, Tokyo, skin. - J. Japan. Soc. Hort. Sci., 64: 463-470. Japan, pp. 760-762. MURATA G., 1971 - , pp. 237-241. - In: KITAMURA YAMAKAWA Y., MORIYA M., ANAMIZU H., 1989 - Char- S., and G. MURATA (eds.) Genshoku-Nippon-no-Shokubut- acterization of hybrid new red-wine grape cultivar ‘Yama suzukan (Mokuhonhen I). Hoikusha, Osaka, Japan. Sauvignon’. - J. Inst. Enol. Vitic. Yamanashi Univ., 24: NAKAGAWA S., 1989 - Studies on collection, preservation and 15-24.

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